Tissue Fragility genes in C. elegans

秀丽隐杆线虫的组织脆性基因

• 批准号: 6849124
• 负责人: JOHN D PLENEFISCH
• 金额: $ 21.6万
• 依托单位: UNIVERSITY OF TOLEDO
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-02-01 至 2009-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6849124
• 关键词: Caenorhabditis elegans; antibody; biomechanics; cell adhesion; cell cycle; cell growth regulation; extracellular matrix proteins; gene expression; genetic models; green fluorescent proteins; helminth genetics; intercellular connection; intermediate filaments; model design /development; molecular cloning; mutant; protein localization; protein quantitation /detection; skin; tissue support frame

DESCRIPTION (provided by applicant): Using C. elegans as a simple in vivo model, the developmental assembly of hemidesmosome (HD) and intermediate filament (IF) organization is being investigated. The epidermis contains a stress resistant network of IFs through which force can be transmitted. Mutations in mua-2, mua-3, mua-5, and mua-6 all result in a growth and use-dependent fragility of this HD/IF mediated trans-epithelial force transmission pathway. Two of these genes, mua-6 and mua-3, have been cloned and encode an IF and a HD associated protein, respectively. The other two genes, mua-5, and mua-2 whose products are as yet unidentified, have been shown to affect the integrity of these same epidermal complexes. These genes may have human homologues that are involved in the etiology of human diseases, but more importantly, the proposed research continues to develop a simple genetic model in which the biology of matrix receptors, cell adhesion, and their control in development can be studied, impacting on our basic understanding of these interactions in normal development and how they may go awry to cause misattachment or attachment failure in muscular dystrophies, blistering skin diseases and cancers. The long term objective is to understand the cell and molecular biology of HD and IF assembly in growth and development. The short term objective is to ascertain the molecular identity of the products of mua-5 and mua-2 and to understand the potential roles played by them in epidermal tissue integrity and the development of HD/IF complexes. The specific aims are: 1) Clone and identify the predicted products of mua-5, and mua-2 using positional rescue, 2) to identify the cellular and sub-cellular localization of MUA-5 and MUA-2 using GFP fusion constructs and antibodies, 3) to examine the localization of HD associated proteins in mua-2 and mua-5, as well as selected other mutants, and the colocalization of these proteins and MUA-2 and MUA-5 using antibodies and GFP reporters.

描述（由申请人提供）：使用秀丽隐杆线虫作为简单的体内模型，正在研究半桥粒（HD）和中间丝（IF）组织的发育组装。表皮包含一个抗压力的中频网络，通过它可以传递力。 mua-2、mua-3、mua-5 和 mua-6 的突变都会导致这种 HD/IF 介导的跨上皮力传递途径的生长和使用依赖性脆弱性。其中两个基因 mua-6 和 mua-3 已被克隆，分别编码 IF 和 HD 相关蛋白。另外两个基因 mua-5 和 mua-2 的产物尚未确定，已被证明会影响这些相同表皮复合体的完整性。这些基因可能具有与人类疾病病因学相关的人类同源物，但更重要的是，拟议的研究继续开发一种简单的遗传模型，在该模型中可以研究基质受体的生物学、细胞粘附及其在发育中的控制，影响我们对正常发育中这些相互作用的基本理解，以及它们如何出错导致肌肉营养不良、起泡性皮肤病和癌症中的错误依恋或依恋失败。长期目标是了解生长和发育过程中 HD 和 IF 组装的细胞和分子生物学。短期目标是确定 mua-5 和 mua-2 产物的分子特性，并了解它们在表皮组织完整性和 HD/IF 复合物发育中发挥的潜在作用。具体目标是：1) 使用定位救援克隆并鉴定 mua-5 和 mua-2 的预测产物，2) 使用 GFP 融合构建体鉴定 MUA-5 和 MUA-2 的细胞和亚细胞定位，以及抗体，3) 使用抗体和 GFP 检查 HD 相关蛋白在 mua-2 和 mua-5 以及选定的其他突变体中的定位，以及这些蛋白与 MUA-2 和 MUA-5 的共定位记者。